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Mortalin/HSPA9 targeting selectively induces KRAS tumor cell death by perturbing mitochondrial membrane permeability

Oncogene volume 39, pages 4257–4270 (2020)Cite this article

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Abstract

The mitochondrial HSP70 chaperone mortalin (HSPA9/GRP75) is often upregulated and mislocalized in MEK/ERK-deregulated tumors. Here, we show that mortalin depletion can selectively induce death of immortalized normal fibroblasts IMR90E1A when combined with K-RasG12V expression, but not with wild-type K-Ras expression, and that K-RasG12V-driven MEK/ERK activity is necessary for this lethality. This cell death was attenuated by knockdown or inhibition of adenine nucleotide translocase (ANT), cyclophilin D (CypD), or mitochondrial Ca2+ uniporter (MCU), which implicates a mitochondria-originated death mechanism. Indeed, mortalin depletion increased mitochondrial membrane permeability and induced cell death in KRAS-mutated human pancreatic ductal adenocarcinoma (PDAC) and colon cancer lines, which were attenuated by knockdown or inhibition of ANT, CypD, or MCU, and occurred independently of TP53 and p21CIP1. Intriguingly, JG-98, an advanced MKT-077 derivative, phenocopied the lethal effects of mortalin depletion in K-RasG12V-expressing IMR90E1A and KRAS-mutated tumor cell lines in vitro. Moreover, JG-231, a JG-98 analog with improved microsomal stability effectively suppressed the xenograft of MIA PaCa-2, a K-RasG12C-expressing human PDAC line, in athymic nude mice. These data demonstrate that oncogenic KRAS activity sensitizes cells to the effects of mortalin depletion, suggesting that mortalin has potential as a selective therapeutic target for KRAS-mutated tumors.

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Fig. 1: Mortalin depletion selectively suppresses K-RafG12V-expressing IMR90E1A cells.
Fig. 2: ANT, CypD and MCU are required for mortalin depletion to induce lethality in K-RasG12V-expressing IMR90E1A cells.
Fig. 3: Mortalin is upregulated in PDAC and its depletion suppresses PDAC cells in vitro.
Fig. 4: Increased mitochondrial permeability is associated with mitochondrial cell death induced by mortalin depletion in MIA PaCa-2 cells.
Fig. 5: TP53 and p21 are not necessary for mortalin depletion to induce mitochondrial cell death.
Fig. 6: MKT-077 derivatives suppress K-Rasmut-expressing IMR90E1A and tumor cells.
Fig. 7: JG-231 suppresses MIA PaCa-2 xenografts in mice.

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Data availability

All data needed to evaluate the conclusions in the paper are present in the paper or the Supplementary Materials. JG series HSP70 modulators are available from JG under a material transfer agreement with UCSF.

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Acknowledgements

We thank Drs Yuri Lazebnik (CSHL), Richard Mulligan (Harvard Univ.), Bert Vogelstein (Johns Hopkins Univ.), and Bob Deschenes (Univ. South Florida) for IMR90E1A, pHAGE, HCT116 cell lines, and the K-RasG12V-encoding gene, respectively. We also thank former Park lab members for technical supports.

Funding

This work was supported by NIH/National Cancer Institute grant (R01CA138441) to JP. Development of HSP70 modulators was funded by NIH R01NS059690 to JG.

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Authors and Affiliations

  1. Department of Biochemistry, Medical College of Wisconsin, Milwaukee, WI, 53226, USA

    Pui-Kei Wu, Seung-Keun Hong, Dmytro Starenki & Jong-In Park

  2. Department of Pathology, Johns Hopkins University, School of Medicine, Baltimore, MD, 21287, USA

    Kiyoko Oshima

  3. Department of Pharmaceutical Chemistry, University of California San Francisco, San Francisco, CA, 94158, USA

    Hao Shao & Jason E. Gestwicki

  4. LaBahn Pancreatic Cancer Program and Department of Surgery, Medical College of Wisconsin, Milwaukee, WI, 53226, USA

    Susan Tsai

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Contributions

Conceptualization, PW and JP; methodology, PW, SH, DS and JP; investigation, PW, SH, DS and JP; formal Analysis, PW, SH, DS, KO and JP; writing—original draft, PW and JP; writing—review and editing, PW, JG, ST and JP; funding acquisition, JP; resources, ST, HS and JG; supervision, JP.

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Correspondence to Jong-In Park.

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JG is an inventor on a US patent application that includes HSP70 modulators. Other authors declare that they have no conflict of interest.

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Wu, PK., Hong, SK., Starenki, D. et al. Mortalin/HSPA9 targeting selectively induces KRAS tumor cell death by perturbing mitochondrial membrane permeability. Oncogene 39, 4257–4270 (2020). https://doi.org/10.1038/s41388-020-1285-5

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